Cellular responses to external mechanical stimuli when seeded to 3D collagen

INTRODUCTION: Collagen is a naturally occurring visco-elastic protein and widely used biomaterial in Tissue engineering. Mechanical stimulation of cell seeded collagen constructs and its effects on cell orientation, intracellular signalling and molecular responses have been reported in literature2. Monitoring of cellular responses to mechanical stimulation include synthesis of active regulatory molecules such as growth factors or hormones, changes in matrix synthesis, cell alignment and enzyme release. The aim of this study was to investigate cellular responses to pre strained, stiffer and more organised collagen bio-artificial matrices

Measurement of collagen synthesis by cells grown under different mechanical stimuli

INTRODUCTION: The use of scaffolds in tissue engineering is essential to provide cells with a matrix for cell proliferation and differentiation resulting in tissue regeneration. Normally this process involves seeding cells onto an artificial biodegradable scaffold providing mechanical support for cells until there is sufficient extracellular matrix deposition (ECM) to replace the artificial scaffold. Collagen is the bulk protein found in the ECM and measurement of its synthesis is the most direct, absolute indicator of ECM production

Durotactic control within a 3D collagen matrix

INTRODUCTION: While matrix stiffness has been implicated in cell adhesion and migration, most studies have focused on the effects of substrate stiffness in 2D. This work describes a novel continuous stiffness gradient model for studying such processes in 3D

Contractile properties of fibroblasts derived from primary frozen shoulder and effects of TGF beta 1 stimulation

INTRODUCTION: Primary Frozen Shoulder (PFS) is a debilitating disease of unknown aetiology. There is fibrosis and contracture of the coracohumeral ligament, tissues of the rotator interval and the glenohumeral ligaments, leading to restrictive shoulder movements requiring surgical intervention [1]. Frozen shoulder has been postulated to be dupuytren’s disease of the shoulder with an association inferred since 1936. The purpose of the study was to test the hypothesis that cellular mechanisms of fibroblasts derived from primary frozen shoulder exhibited similar activity in terms of contraction and response to cytokine (transforming growth factor beta1) to fibroblasts derived from dupuytren’s disease. Understanding of cellular responses is critical to developing non surgical treatment strategies

Profile of pulmonary smooth muscle cells and their response to blockade and Stimulations

INTRODUCTION: The tone of the pulmonary arteries is the summation of the activity of each smooth muscle cell (SMC) within a vessel wall and its interaction with the endothelial cells and extracellular matrix (including collagen). There are reported phenotypic differences between SMC in the inner & outer layers of pulmonary artery walls1. To tissue engineer a blood vessel a basic understanding of differences between inner and outer SMC’s in terms of attachment and contraction of 3D matrix is essential. We hypothesised that they will differ in their ability to contract a 3D collagen gel. Using a Culture Force Monitor (CFM) we sought to: (1) quantify the contractile ability of SMC derived from inner and outer normal and hypoxic arteries, harvested from piglet models, over 24 hours, (2) test the effect of cytoskeletal blockade on force retention in collagen gels and (3) quantify cellular contractile response to agonist or antagonist stimulation

External loading determines specific ECM genes regulation

Bio artificial matrices embedded with cells are simulated in bioreactors to facilitate ECM production. As cells attach, they develop forces, which are dependent on cell type and matrix stiffness. External forces (i.e strain), however, are critical for tissue homeostasis and elicit specific cellular responses, such as gene expression and protein production. Collagen Type I is a widely used scaffold in Tissue engineering. The aim of this study was to study the mechanical and molecular responses, of different cell types to increasing collagen substrate stiffness

Pulmonary artery smooth muscle cells response to vaso-active stimulations in a real-time 3-Dimensional model

INTRODUCTION: The tone of the pulmonary arteries is the summation of the activity of each smooth muscle cell (PASMC) within a vessel wall and its interaction with the endothelial cells and extracellular matrix (including collagen). There are reported phenotypic differences between PASMC in the inner & outer layers of pulmonary artery walls1. The response of a tissue engineered blood vessel to contractile and relaxing stimulants in-vitro is essential to predicting the response of the physiological and pathological vessels in-vivo. Previous work showed that pulmonary artery relaxation to nitric oxide is inhibited after exposure to chronic hypoxia 2. We hypothesised that PASMC will differ in their ability to contract or relax a 3D collagen gel. Using a Culture Force Monitor (CFM) we sought to quantify the cellular response of PASMC derived from inner and outer normal and hypoxic arteries, harvested from piglet models, over 24 hours in response to contractile agonists and relaxing antagonists

Innervation and Muscle Cell Infiltration of Plastic Compressed Collagen Constructs

Abstract not available

A Novel Tenorrhaphy Suture Technique with Tissue Engineered Collagen Graft to Repair Large Tendon Defects

Surgical management of large tendon defects with tendon grafts is challenging, as there are a finite number of sites where donors can be readily identified and used. Currently, this gap is filled with tendon auto-, allo-, xeno-, or artificial grafts, but clinical methods to secure them are not necessarily translatable to animals because of the scale. In order to evaluate new biomaterials or study a tendon graft made up of collagen type 1, we have developed a modified suture technique to help maintain the engineered tendon in alignment with the tendon ends. Mechanical properties of these grafts are inferior to the native tendon. To incorporate engineered tendon into clinically relevant models of loaded repair, a strategy was adopted to offload the tissue engineered tendon graft and allow for the maturation and integration of the engineered tendon in vivo until a mechanically sound neo-tendon was formed. We describe this technique using incorporation of the collagen type 1 tissue engineered tendon construct

Generating intrafusal skeletal muscle fibres in vitro: Current state of the art and future challenges

Intrafusal fibres are a specialised cell population in skeletal muscle, found within the muscle spindle. These fibres have a mechano-sensory capacity, forming part of the monosynaptic stretch-reflex arc, a key component responsible for proprioceptive function. Impairment of proprioception and associated dysfunction of the muscle spindle is linked with many neuromuscular diseases. Research to-date has largely been undertaken in vivo or using ex vivo preparations. These studies have provided a foundation for our understanding of muscle spindle physiology, however, the cellular and molecular mechanisms which underpin physiological changes are yet to be fully elucidated. Therefrom, the use of in vitro models has been proposed, whereby intrafusal fibres can be generated de novo. Although there has been progress, it is predominantly a developing and evolving area of research. This narrative review presents the current state of art in this area and proposes the direction of future work, with the aim of providing novel pre-clinical and clinical applications